Effect of macrophage polarization M1 or M2 subtypes on iron metabolism
Objective To explore the effect of macrophage polarization M1 or M2 subtypes on iron metabolism.Methods Macrophages(RAW264.7)were treated with lipopolysaccharide(LPS)20 ng/mL or IL-4 10 ng/mL in cell culture.The morphological characteristics of polarized macrophages were observed under a light microscope.The expression levels of M1-type markers TNF-α,inducible nitric oxide synthase(iNOS)and M2-type marker arginase 1(Arg-1)in macrophages were detected with immunofluorescence staining and Western blot.LPS or IL-4 treated macrophages were co-cultured with erythrocyte or iron;the iron intake of macrophages was evaluated by Wright's staining and iron staining,ferritin levels in macrophages and culture medium were determined using ELISA assay.Results After LPS treatment,the microscopic appearance of macrophages was changed from circular to polymorphic shapes;the expression levels of TNF-α and iNOS were significantly increased;and macrophages exhibited erythrocytophagy with a significant increase of iron particles in the cytoplasm.After IL-4 treatment,the morphology of macrophages also showed polymorphic changes,and the expression of Arg-1 was increased;however,no erythrocytophagy was observed,and there were fewer iron particles phagocytosed in the cytoplasm.In addition,the intracellular ferritin levels of macrophages treated with LPS were significantly increased compared to the IL-4 treatment group.Conclusion LPS and IL-4 can induce macrophage polarization into different subtypes,LPS-induced M1 macrophages have an increased uptake and storage of iron compared to IL-4-induced M2 macrophages.
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